WO2020181825A1

WO2020181825A1 - Nucleic acid extraction apparatus

Info

Publication number: WO2020181825A1
Application number: PCT/CN2019/119366
Authority: WO
Inventors: 陈辉
Original assignee: 陈辉
Priority date: 2019-03-12
Filing date: 2019-11-19
Publication date: 2020-09-17
Also published as: JP2022520532A; EP3940052A1; US20220119799A1; EP3940052A4

Abstract

Disclosed is a nucleic acid extraction apparatus. The nucleic acid extraction apparatus comprises a workbench having a table top, and further comprises a sliding seat, a sample rack, a reagent strip rack, a pipetting table, a piston assembly and a suction assembly. The nucleic acid extraction apparatus comprises the sliding seat and the pipetting table, the sample rack and the reagent strip rack are arranged on the sliding seat, the piston assembly and the suction assembly are respectively arranged on the pipetting table, the sample rack is provided with sample holes, the reagent strip rack is provided with accommodation slots, the number of accommodation slots corresponds to that of the sample holes on a one-to-one basis, and when reagent strips are respectively accommodated in the accommodation slots, a first suction head hole, second suction head holes and various reagent holes in each reagent strip respectively correspond to the corresponding sample holes, such that the loading of a suction head assembly and suction heads and operations such as suction, injection and mixing of a sample and various reagents can be realized by means of a vertical movement, relative to the sliding seat, of the pipetting table and a horizontal movement, relative to the pipetting table, of the sliding seat, thereby realizing the automated operation of nucleic acid extraction.

Description

Nucleic acid extraction equipment

This application requires that the applicant Chen Hui filed a Chinese invention patent application with the application number 201910184157.7 and the title of "a nucleic acid extraction device" with the Chinese Patent Office on March 12, 2019, and the applicant filed with the Chinese Patent Office on the same filing date. The priority of the Chinese utility model patent application filed with the application number 201920311623.9 and the invention title is "a nucleic acid extraction device", the entire content of the application is incorporated herein by reference.

Technical field

The present invention relates to the field of biotechnology, in particular to a nucleic acid extraction device.

Background technique

Nucleic acid extraction is a very important technology in life science research, biotechnology application and genetic diagnosis. Early nucleic acid extraction was based on the principle of phenol-chloroform extraction to remove impurities and ethanol precipitation of nucleic acids, but this method is very cumbersome.

At present, commercial nucleic acid extraction methods are mainly based on solid phase adsorption methods, that is, nucleic acid molecules are adsorbed on the surface of a solid phase adsorbent under specific conditions (such as pH, specific salts, etc.), and then eluted. Purification methods mainly include the following two: (1) Spin column method: Fill the spin column with silicon-containing materials or other solids that can absorb nucleic acids as a medium, and use chaotropic salts such as guanidine salts as binding agents. When nucleic acid molecules pass When the spin column is adsorbed on the solid phase, the impurities are washed away with a solution containing ethanol, and finally the nucleic acid is eluted with a low-salt solution; this method is simple and convenient, with high extraction efficiency and good extraction effect, which is currently the most used The disadvantage of the nucleic acid purification method is that it requires repeated centrifugation during the extraction process, which is not convenient for high-throughput and automated operations. (2) Magnetic bead method: Use magnetic microspheres coated with silicon materials or other materials that can bind nucleic acids as solid phase carriers, under the action of chaotropic salts (guanidine hydrochloride, guanidine isothiocyanate, etc.) and an external magnetic field , Separate DNA and RNA from samples; this method is easy to realize automation and high-throughput operation, but the disadvantage is that the cost is high and the extraction process is time-consuming, which leads to the long time consuming of the entire genetic test, and doctors and patients cannot obtain test results in time. The magnetic beads have poor chemical stability. The extraction efficiency is easily affected, and the extraction effect of magnetic beads from different samples, manufacturers, and batches varies greatly. Lead to unstable extraction and purification results. In addition, the repeated washing of the magnetic beads during the magnetic bead extraction process is also prone to cross-contamination, resulting in false positive results. Existing nucleic acid automatic extraction devices are generally based on the above-mentioned magnetic bead method, such as the Chinese invention patent with the patent number ZL201620294747.7 (authorized announcement number CN205635634U), and the Chinese invention with patent number ZL201610687392.2 (authorized announcement number CN106119082A) Nucleic acid extraction device disclosed in patents.

Summary of the invention

The first technical problem to be solved by the present invention is to provide a nucleic acid extraction device with simple structure and convenient use for the prior art.

The second technical problem to be solved by the present invention is to provide a nucleic acid extraction device with high nucleic acid extraction purity in view of the prior art.

The third technical problem to be solved by the present invention is to provide a nucleic acid extraction device that does not cause cross-contamination between samples in the nucleic acid extraction process based on the prior art.

The technical solution adopted by the present invention to solve the above technical problems is: a nucleic acid extraction device, including a workbench with a countertop, characterized in that it further includes:

The sliding seat is arranged on the table, and it can slide back and forth along the table under the drive of the sliding component;

The sample holder is arranged on the sliding seat, and the sample holder is provided with a plurality of sample holes arranged side by side in a direction perpendicular to the sliding direction of the sliding seat, and the sample holes are used for placing the sample tube;

The reagent strip holder is set on the sliding seat, and is provided with a containing groove for placing the reagent strip, and the number of the containing groove matches the number of the sample holes,

Each reagent strip includes a first tip hole, a second tip hole, and a reagent hole that are arranged in a straight line, wherein the first tip hole is used to place the suction head assembly, and the second tip hole is used to place the tip. The reagent hole includes a first reagent hole for containing diluent, a second reagent hole for containing cleaning solution, a third reagent hole for containing eluent, and a fourth reagent for storing target substance There are at least two second reagent holes, each containing slot is placed in the state of the reagent strip, the first tip hole, the second tip hole and each reagent hole on each reagent strip are opposite to the corresponding sample hole ；

The pipetting table is set horizontally on a stand fixed on the table top and can be moved up and down relative to the sliding seat driven by the lifting assembly;

The piston assembly is set on the pipetting table and includes a piston part and a piston driving device. The number of piston parts matches the number of sample holes. Each piston part includes a piston tube and is built into the piston tube and can follow the inner wall of the piston tube The pistons move up and down, and each piston tube is vertically pierced on the pipetting table. The lower end of the piston tube is respectively formed with a first joint for inserting and connecting the suction head. The piston driving device can drive the piston in each piston to correspond to the corresponding The piston tube moves up and down; and

The suction assembly includes a connection suction tube and a vacuum pump. The number of the connection suction tube matches the sample hole, and each connection suction tube is pierced on the pipetting table, and the upper end of each connection suction tube is respectively connected with the vacuum pump,

In order to conveniently heat and heat the sample according to the operation requirements, a heating device for heating the sample holder is also provided on the sliding seat.

In order to easily pierce the seal on the reagent strip, it also includes a piercing component for piercing the seal on each reagent hole. The piercing component includes a piercing frame and a piercing head installed on the piercing frame. The broken head is matched with the sample hole, the pistons are fixed on the piston frame, the piston frame is fixed on the output shaft of the piston driving device, and the piercing frame is fixed with the piston frame. In this way, when the piston driving device drives the piston frame to move down, it drives the piston down and at the same time drives the piercing frame down, so that the piercing heads can move down to pierce the seal on the reagent strip.

In order to better realize the suction action, there are multiple vacuum pumps and correspond to the piston parts one-to-one, the piston frame is provided with suction joints, and the number of the suction joints corresponds to the piston parts one-to-one, and On the piston frame through which each suction joint is inserted, one end of each suction connector is connected with the corresponding interface on the vacuum pump through a connecting pipe, and the other end of the suction connector is connected with the upper end of the suction pipe through a connecting pipe.

In order to enable the vacuum pumps to be installed more stably, a mounting frame is arranged on the table, and the vacuum pumps are respectively placed on the mounting frame.

In order to better realize the sliding movement of the sliding seat along the table surface driven by the sliding assembly, the sliding assembly includes a first sliding block, a first sliding rail, a pressing block, a sliding motor, a timing belt, and a driving pulley And the driven pulley, the sliding block is fixed at the bottom of the sliding seat, the first sliding rail is fixed on the table, the first sliding block can slide back and forth along the first sliding rail, the sliding motor is fixed on the table, and the driving belt The wheel is fixed on the output shaft of the sliding motor, the driven pulley is installed on the table, the timing belt is tensioned between the driving pulley and the driven pulley, and the pressure block is fixed on one side of the sliding seat and pressed on On the synchronous belt.

Preferably, the reagent strip includes a carrier plate, the carrier plate is provided with a plurality of first through holes and second through holes in parallel along its length, and one side of the carrier plate protrudes from the first through hole. Corresponding tube body, and at the same time, one side of the carrier plate also has a sleeve body corresponding to the second through hole, and the nozzle of each tube body is directly opposite to the corresponding first through hole. The openings are respectively opposite to the corresponding second through holes, thereby respectively forming a first tip hole, a second tip hole and each reagent hole.

Further, a first connecting piece and a second connecting piece are respectively provided on both sides of the carrier board, and the first connecting piece and the second connecting piece can be connected in a detachable manner.

Still further, the first connecting piece is a connecting column, and the second connecting piece is a connecting sleeve into which the connecting column is inserted.

Preferably, the suction head assembly includes a filter element suction head and a disposable capillary pipette, the lower end of the filter element suction head is embedded with an adsorption filter element, and the upper end of the disposable capillary pipette is sleeved on the lower end of the connecting straw and is embedded in In the upper port of the filter tip, and the disposable capillary pipette is located in the filter tip and above the adsorption filter. By setting the disposable capillary straw, the connecting straw and the filter tip are separated by the uncontaminated disposable capillary straw to avoid contact between the connecting straw and the filter tip during the suction process, thereby avoiding contamination of the filter tip, and The purity of nucleic acid extraction is ensured. In addition, the connecting pipette can be reused by setting a disposable capillary pipette, thereby reducing the cost of nucleic acid extraction. In addition, the disposable capillary tube allows the solution to be quickly and completely drawn from the filter element through the capillary effect, thereby ensuring the complete separation of nucleic acid and liquid adsorbed on the filter element and the thorough cleaning of impurities on the filter element.

In order to achieve better suction, the lower end of the disposable capillary straw is opposite to the adsorption filter element up and down.

In order to improve the suction effect, the distance between the lower end of the disposable capillary tube and the upper surface of the adsorption filter element is 0-10 mm.

Preferably, the distance between the lower end of the disposable capillary straw and the upper surface of the adsorption filter element is 2-5 mm, so that sufficient suction can be achieved.

Preferably, the disposable capillary straw includes a first part located at the upper end and a second part located at the lower end, wherein the first part is used for sleeve connection on the connecting straw, and the diameter of the second part can supply liquid in Pass the disposable capillary straw smoothly under the action of suction. If the diameter of the second part is too large, first, no capillary action is not conducive to complete liquid extraction, and second, part of the liquid will remain between the second part and the suction tube of the filter element; on the contrary, if the second part is too small, The liquid may be blocked in the second part, and smooth suction cannot be achieved.

Further, preferably, the pipe diameter of the second part is 0.5 to 2.0 mm.

Preferably, the edge of the upper port of the disposable capillary pipette is lower than the edge of the upper port of the filter tip. Through the above design, the second suction head can be realized. The first suction head can separate the filter tip from the disposable capillary straw, and the second suction head can separate the disposable capillary straw from the connecting straw. The filter tip can be put on other tips to clean the adsorption filter in the filter tip, so as to achieve the elution of nucleic acid substances.

In order to facilitate the detachment between the disposable capillary straw and the filter tip, a convex ring is provided along the circumferential direction on the inner peripheral surface of the upper end of the filter tip, and the convex ring abuts against the corresponding outer peripheral surface of the disposable capillary straw. Compared with the surface-to-surface contact between the inner peripheral surface of the disposable capillary straw and the outer peripheral surface of the connecting straw, the frictional force of the linear surface contact between the convex ring and the outer peripheral surface of the disposable capillary straw is smaller. This design makes The filter tip and the disposable capillary straw can be withdrawn from the connecting straw separately. First, the filter tip with less friction is returned, and then the disposable capillary straw, and the setting of the convex ring can make the disposable capillary straw and A seal is formed between the filter tips. In addition, the convex ring can also prevent the adsorption filter from falling from the filter tips.

In order to form a better seal between the disposable capillary straw and the suction head of the filter element, the convex ring is at least two arranged up and down.

In order to further promote the sealing between the disposable capillary straw and the suction head of the filter element, the convex rings are arranged up and down at intervals.

Compared with the prior art, the advantage of the present invention is that the nucleic acid extraction device of the present invention includes a sliding seat and a pipetting table, the sliding seat is provided with a sample holder and a reagent strip holder, and the pipetting table is respectively provided with a piston The component and the suction component, the sample holder has sample holes, and the reagent strip holder has accommodating grooves. The number of the accommodating grooves corresponds to the sample holes one-to-one, and when each accommodating groove contains a reagent strip In the state, the first tip hole, the second tip hole, and each reagent hole on each reagent strip are opposite to the corresponding sample hole, so that the pipetting table moves up and down relative to the sliding seat or the sliding seat is relative to The lateral movement of the pipetting table can realize the loading of the suction head assembly and the suction head, as well as the suction, injection and mixing of samples and various reagents, thereby providing a basis for the automated operation of nucleic acid extraction. In addition, by providing a disposable capillary pipette in the present invention, it is possible to prevent the connecting pipette from contacting the lower end of the connecting pipette during the suction process and contaminating the connecting pipette, thereby ensuring that there will be no pollution between different nucleic acid extraction samples, and connecting the pipette Repeated use.

Description of the drawings

Figure 1 is a schematic structural diagram of a nucleic acid extraction device in an embodiment of the present invention;

Fig. 2 is a schematic structural diagram in another direction of Fig. 1;

3 is a schematic diagram of a partial structure of a nucleic acid extraction device in an embodiment of the present invention;

Figure 4 is an enlarged view of part I in Figure 3;

FIG. 5 is a schematic structural diagram in another direction of FIG. 3;

FIG. 6 is a schematic structural diagram in another direction of FIG. 3;

Figure 7 is a schematic diagram of another partial structure of a nucleic acid extraction device in an embodiment of the present invention;

FIG. 8 is still another partial structural diagram of the nucleic acid extraction device in the embodiment of the present invention;

Figure 9 is a schematic diagram of the structure of the head back plate in the embodiment of the present invention;

10 is a schematic diagram of the structure of the reagent strip holder in the embodiment of the present invention;

11 is a schematic diagram of the structure of the sample holder in the embodiment of the present invention;

Figure 12 is an exploded view of the structure of the piston in the embodiment of the present invention;

Figure 13 is a schematic diagram of the structure of a reagent strip in an embodiment of the present invention;

FIG. 14 is a schematic structural diagram in another direction of FIG. 13;

FIG. 15 is a schematic structural diagram in still another direction of FIG. 13;

Figure 16 is a schematic structural view of the suction head assembly in the embodiment of the present invention;

Figure 17 is a cross-sectional view of the suction head assembly in the embodiment of the present invention;

Figure 18 is an enlarged view of part Ⅱ in Figure 17;

Figure 19 is a schematic diagram of the structure of the filter tip in the embodiment of the present invention;

Figure 20 is an exploded view of the structure of the suction head assembly in the embodiment of the present invention.

detailed description

The present invention will be further described in detail below in conjunction with the embodiments of the drawings.

As shown in Figures 1-20, a nucleic acid extraction device includes a worktable 1 with a table top 10, a sliding seat 21, a sample holder 3, a reagent strip holder 213, a pipetting table 4, a piston assembly, and a suction assembly. In this embodiment, a housing 11 is provided on the table top 10, and the sliding seat 21 and the pipetting table 4 are both provided in the housing 11. In addition, a heat dissipation fan 13 is provided on the back of the housing 11.

The sliding seat 21 is arranged on the table 10 and can slide back and forth along the table 10 under the drive of the sliding assembly 22. In this embodiment, in order to better realize the sliding movement of the sliding seat 21 along the table 10 driven by the sliding assembly 22, the sliding assembly 22 includes a first sliding block 227, a first sliding rail 226, and a pressing block 225 , The sliding motor 221, the timing belt 224, the driving pulley 222 and the driven pulley 223, the first sliding block 227 is fixed at the bottom of the sliding seat 21, the first sliding rail 226 is fixed on the table 10, the first The sliding block 227 can slide back and forth along the first sliding rail 226, the sliding motor 221 is fixed on the table 10, the driving pulley 222 is fixed on the output shaft of the sliding motor 221, and the driven pulley 223 is installed on the table 10 , The timing belt 224 is tensioned between the driving pulley 222 and the driven pulley 223, and the pressure block 225 is fixed on one side of the sliding seat 21 and pressed on the timing belt 224. In this embodiment, the sliding motor 221 is a stepping motor.

The sample holder 3 is arranged on the sliding seat 21, and the sample holder 3 has a plurality of sample holes 31 arranged in a direction perpendicular to the sliding direction of the sliding seat 21, and the sample holes 31 are used to place the sample tube 15 . In this embodiment, the sample holder 3 is in the shape of a long block, and the above-mentioned sample holes 31 are evenly distributed along the length direction, and each sample hole 31 extends along the height direction of the sample holder 3 to form a blind hole shape. In order to conveniently heat and keep the sample according to operation requirements, a heating device for heating the sample tube 15 placed in the sample hole 31 is also provided on the sliding seat 21. Specifically, in this embodiment, the heating device is a heating bath 211 provided at the front end of the sliding seat 21, and the sample holder 3 is contained in the heating bath 211, so that the sample on the sample holder 3 can be fully heated and kept warm. .

The reagent strip holder 213 is arranged at the rear end of the sliding seat 21, on which is provided a containing groove 213a for placing the reagent strip 7, and the number of the reagent strip 7 containing grooves 213a corresponds to the sample hole 31 one-to-one The above reagent strips 7 all include a first tip hole 71, a

second tip hole

72a, 72b and a reagent hole that are arranged in a straight line. The first tip hole 71 is used to place the suction head assembly, and the

second tip hole

72a , 72b is used to place the tip 82, the reagent hole includes a first reagent hole 73 for containing the diluent, a

second reagent hole

74a, 74b, 74c, 74d for containing the cleaning solution, and for containing the elution The third reagent hole 75 of the liquid and the fourth reagent hole 76 for storing the target substance, each of the accommodating grooves 213a is in the state where the reagent strip 7 is placed, and the first tip hole 71 and the second tip hole 71 on each reagent strip 7 The two

tip holes

72a, 72b and each reagent hole are respectively opposite to the corresponding sample hole 31. In this embodiment, the above-mentioned reagent strip 7 includes a carrier plate 70. The carrier plate 70 is provided with a plurality of first through holes 701 and second through holes 702 side by side along its length. One side of the carrier plate 70 extends The tube body 703 corresponding to the first through hole 701 is one-to-one, and at the same time, a sleeve body 704 corresponding to the second through hole 702 is extended on one side of the carrier plate 70, and the nozzle of each tube body 703 Respectively face the corresponding first through holes 701 and the openings of each sleeve 704 face the corresponding second through holes 702 respectively, thereby respectively forming the aforementioned first nozzle hole 71,

second nozzle hole

72a, 72b and Each reagent well. Further, the

second reagent holes

74a, 74b, 74c, 74d contain cleaning liquids, which are the first cleaning liquid, the second cleaning liquid, the third cleaning liquid, and the fourth cleaning liquid, respectively, so that during the nucleic acid extraction process, Repeated rinsing can effectively ensure the quality of nucleic acid extraction.

Furthermore, a first connecting member 78 and a second connecting member 77 are respectively provided on both sides of the above-mentioned carrier plate 70, and the first connecting member 78 and the second connecting member 77 can be connected in a detachable manner. In this way, two or more reagent strips 7 can be connected by the above-mentioned first connecting member 78 and second connecting member 77, so that the connected and combined reagent strip 7 can be placed upright without additional devices to realize the reagent strip. 7 is placed to facilitate the use of reagent strip 7. The detachable connection of the first connecting member 78 and the second connecting member 77 can be realized in a variety of specific ways. Preferably, the first connecting member 78 is a connecting column, and the second connecting member 77 is for inserting the connecting column. Connection sleeve. Specifically, the first connecting member 78 is fixed to the bottom of the side edge of the carrier 70, and the first connecting member 78 extends vertically downward and is suspended, and the second connecting member 77 is fixed to the other of the carrier 70. The bottom of one side edge and/or the corresponding tube body 703 is at the corresponding tube wall of the side, and the second connecting member 77 extends vertically downward and its opening is exposed outside.

The pipetting table 4 is horizontally arranged on a stand 12 fixed to the table 10, and can be moved up and down relative to the sliding seat 21 under the driving of the lifting assembly 64. In order to better realize the up and down lifting of the liquid transfer table 4 driven by the lifting component 64, the lifting component 64 includes a lifting motor 641, a lifting screw 642, and a lifting seat 643. The above stand 12 includes a door frame 121 and fixed at The vertical plate 122 on the top plate of the door frame 121, the above-mentioned lifting motor 641 is installed at the upper end of the vertical plate 122, and the vertical plate 122 is respectively provided with a second sliding rail 122b and sliding holes 122a extending vertically below the lifting motor 641 The lifting screw 642 is vertically arranged and its upper end is connected to the output shaft of the lifting motor 641, and the lower end is connected to the top plate of the door frame 121. The lifting seat 643 is penetrated on the vertical plate 122 through the sliding hole 122a, and its The front end is fixed with the above-mentioned pipetting table 4, the rear end is sleeved on the above-mentioned screw rod 642 and forms a screw transmission pair with the screw rod 642, and the lifting seat 643 is provided with a second sliding block 644, and the above-mentioned lifting motor 641 drives the wire When the rod 642 rotates, the screw rod 642 can drive the lifting base 643, and the second sliding block 644 can move up and down along the second sliding rail 122b, and the corresponding part of the lifting base 643 can move up and down along the sliding hole 122a. In the initial state, the reagent strip holder 231 is located below the door frame 121.

The piston assembly is arranged on the pipetting table 4 and includes a piston member 62 and a piston driving device 61. In this embodiment, the piston driving device 61 is a motor, and the number of the piston members 62 is the same as the number of the sample holes 31 Correspondingly, each piston member 62 includes a piston tube 622 and a piston 621 built in the piston tube 622 and capable of moving up and down along the inner wall of the piston tube 622, and each piston tube 622 is vertically pierced on the pipetting table 4, the piston The lower end of the tube 622 is respectively formed with a first joint 621a for inserting the suction head 82, and the piston driving device 61 can drive the piston 621 in each piston member 62 to move up and down relative to the corresponding piston tube 622. In this embodiment, the piston driving device 61 is a through-type stepping motor, the pistons 621 are fixed on the piston frame 63, and each piston 621 includes a piston body 6210 and a piston rod 6211 fixed on the top end of the piston body 6210. The piston frame 63 is fixed on the output shaft of the piston driving device 61. In order to make the installation of the vacuum pumps 5 more stable, a mounting frame 51 is provided on the table 10, and the vacuum pumps 5 are placed on the mounting frame 51 respectively.

The above-mentioned suction assembly includes a connecting suction tube 91 and a vacuum pump 5. The number of the connecting suction tube 91 corresponds to the above-mentioned sample hole 31 one-to-one, and each connecting suction tube 91 is pierced on the pipetting table 4, and each connected to the upper end of the suction tube 91 They are respectively connected to the vacuum pump 5, and the lower ends are respectively formed with second joints 911 for inserting and connecting the above-mentioned suction head assembly. In order to better realize the suction action, there are multiple vacuum pumps 5 and correspond to the connecting suction pipes 91 one-to-one. The piston frame 63 is provided with suction joints 92. The number of the suction joints 92 and the connection suction pipes 91 There is a one-to-one correspondence, and each suction joint 92 is threaded on the suction joint frame 65. One end of each suction joint 92 is connected to the corresponding port on the vacuum pump 5 through a connecting pipe (not shown), and the other end is connected The upper end interface of the connecting straw 91 is connected through a connecting tube (not shown).

The suction head assembly includes a filter suction head 83 and a disposable capillary suction tube 81. The lower port of the connection suction tube 91 is inserted into the upper end of the filter suction head 83, and the lower end of the filter suction head 83 is embedded with an adsorption filter 830. The upper end of the disposable capillary straw 81 is sleeved on the lower end of the connecting straw 91 and embedded in the upper port of the filter tip 83, and the disposable capillary straw 81 is located in the filter tip 83 and in the adsorption filter 830 The lower end of the disposable capillary pipe 81 is opposite to the adsorption filter element 830 up and down. In this way, in this embodiment, the disposable capillary pipe 81 can prevent the connecting pipe 91 from contacting the filter tip 83 during the suction process, thereby avoiding the filter tip 83 from being contaminated, thereby ensuring the purity of nucleic acid extraction.

Furthermore, the lower end of the disposable capillary tube 81 is opposite to the adsorption filter element 830 up and down. In order to maximize the suction effect, the lower end of the disposable capillary tube 81 and the upper surface of the adsorption filter element 830 maintain a proper distance. In this embodiment, in order to achieve sufficient suction, the distance between the lower end of the disposable capillary straw 81 and the upper surface of the adsorption filter element 830 is 4 mm.

The disposable capillary straw 81 includes a first portion 811 located at the upper end and a second portion 812 located at the lower end. The first portion 811 is used to sleeve the connecting straw 91, and the diameter of the second portion 812 can be The supply liquid smoothly passes through the disposable capillary tube 81 under the action of suction. If the tube diameter of the second part 812 is too large, the capillary action of the second part 812 is not significant, and the liquid will remain between the second part 812 and the connecting straw 91. On the contrary, if the tube diameter of the second part 812 is too small , The liquid will be blocked in the second part 812, and smooth suction cannot be achieved. In this embodiment, preferably, the pipe diameter of the second part 812 is 0.5 to 2.0 mm.

Still further, in order to realize the secondary suction head, the mouth edge of the upper port of the disposable capillary straw 81 is lower than the mouth edge of the upper port of the above-mentioned filter tip 83. In this way, the first withdrawal of the tip makes the disposable capillary tube 81 and the filter tip 83 separate, and the second withdrawal of the tip can make the disposable capillary tube 81 and the connecting straw 91 separate, so that the filter tip 83 can be covered after the tip is withdrawn. Go to other tips to clean the adsorption filter 830 in the filter tip 83 to achieve the elution of nucleic acid substances.

In addition, in order to facilitate the separation between the disposable capillary straw 81 and the filter tip 83, the inner peripheral surface of the upper end of the filter tip 83 is provided with a convex ring 831 along the circumferential direction, and the convex ring 831 is connected to the disposable capillary straw 81 The outer peripheral surfaces of the corresponding parts are offset. Compared with the surface-to-surface contact between the outer peripheral surface of the disposable capillary straw 81 and the inner peripheral surface of the filter tip 83, the frictional force of the linear surface contact between the convex ring 831 and the outer peripheral surface of the disposable capillary straw 81 is smaller This design allows the filter tip 83 and the disposable capillary straw 81 to be withdrawn from the connecting straw 91 respectively. First, the filter tip 83 with lower friction is removed, and then the disposable capillary tube 81, and the configuration of the convex ring 831 can make the disposable capillary tube 81 and the filter tip 83 form a seal. In this embodiment, in order to form a better seal between the disposable capillary straw 81 and the filter tip 83, the above-mentioned raised rings 831 are two arranged up and down, and the two raised rings 831 are arranged up and down to be further The sealing between the secondary tube and the filter tip 83 is promoted. In addition, the design of the convex ring 831 can also prevent the adsorption filter 830 from falling off the filter tip 83.

In order to easily pierce the seal on the reagent strip 7, the nucleic acid extraction device of the present invention further includes a piercing assembly for piercing the seal on each reagent hole. The piercing assembly includes a piercing rack 141 and a The piercing head 14 on the piercing frame 141 corresponds to the sample hole 31 one-to-one, and the piercing frame 141 and the piston frame 63 are fixed as a whole. In this way, when the lifting motor 641 drives the liquid transfer table 4 to move down, and the piston drive device 61 drives the piston frame 63 to move down and drives the piston 621 to move down, the piercing frame 141 is driven to move down, and the piercing heads 14 can be moved down. And pierce the seal on each reagent hole of reagent strip 7. In order to facilitate the unloading of the suction head 82 on the first joint 621a and the suction head assembly on the second joint 911, an ejector plate 41 is fixed at the bottom of the pipetting table 4, and the ejector plate 41 is respectively provided with a first ejector head. Hole 41a and second back-out hole 41b, the first joint 621a and second joint 911 are respectively penetrated in the first back-out hole 41a and second back-out hole 41b, and the aperture of the first back-out hole 41a is the same as The corresponding radius of the first joint 621a matches and is smaller than the radius of the connection end of the filter tip 83, and the aperture of the second head-out hole 41b matches the radius of the second joint 911 and is smaller than the radius of the connection end of the suction head 82. Two ends of the pipetting table 4 are respectively vertically provided with guide posts 411. When the piston driving device 61 drives the piston frame 63 to move down to a certain distance, the piston frame 63 can abut against the top of the guide posts 411. Each guide post 411 is pierced on the pipetting table 4 and can move up and down relative to the pipetting table 4, and its lower ends are respectively fixed to the above-mentioned head-ejection plate 41, and each guide post 411 has a structure with both ends thick and thin in the middle. In addition, a return spring 411a is penetrated in the middle of each guide post 411. Specifically, the lower end of each guide post 411 is inserted into the pipetting table 4, and the return spring 411a is limited between the upper end of the guide post 411 and the top surface of the pipetting table 4. When the piston driving device 61 drives the piston frame When 63 moves down to a certain distance, the piston frame 63 abuts against the top of each guide post 411, thereby driving the head ejection plate 41 to move down to realize the suction head assembly or suction head 82. When the pressure on each guide post 411 When it disappears, each guide post 411 is reset under the action of the corresponding return spring 411a. In this way, by providing the head-back plate 41, the suction head 82 can be unloaded from the first joint 621a, or the suction head assembly can be unloaded from the second joint 911, or the filter cartridge suction head 83 can be removed from the disposable capillary pipe 81 Uninstall on.

The working process of the present invention is as follows:

Put the sample tube 15 and the reagent strip 7 into the sample holder 3 and the reagent strip holder 231 respectively, start the nucleic acid extraction device, reset each motor, and the heating device heats and keeps the sample according to the set heat preservation program. After the sample is kept warm, the piston driving device 61 cooperates with the lifting motor 641 to drive the piercing frame 141, so that each piercing head 14 pierces the seal on the corresponding reagent hole, and is ready for subsequent operations.

The sliding motor 221 drives the sliding seat 21 so that the suction heads 82 on the sliding seat 21 are directly opposite to the first joints 621a on the pipetting table 4, and the lifting motor 641 drives the pipetting table 4 to move down. The first joint 621a presses the suction head 82 and rises. The sliding motor 221 drives the sliding seat 21 so that each first reagent hole 73 containing the diluent is directly opposite to each suction head 82, the pipetting table 4 is lowered, and each suction head 82 is inserted into the corresponding first reagent. In the hole 73, the piston driving device 61 works, and the suction head 82 sucks the diluent. After the suction is completed, the lifting motor 641 drives the pipetting table 4 to rise.

The sliding seat 21 translates so that the sample tubes 15 on the sample holder 3 and the first joints 621a on the pipetting table 4 are aligned one by one, the lifting motor 641 drives the pipetting table 4 to move down, and the piston driving device 61 works. The diluent in each tip 82 is added to the corresponding sample tube 15, and the piston member 62 is sucked several times to make the diluent and the sample fully mixed, and keep it warm for 5-10 minutes as needed.

The lifting motor 641 drives the pipetting table 4 to rise, the sliding motor 221 aligns the

second nozzle holes

72a, 72b with the first joints 621a one by one, the pipetting table 4 descends, the piston driving device 61 works, and drives the head back The plate 41 retracts each suction head 82 into the corresponding second

suction head hole

72a, 72b. The pipetting table 4 rises, the sliding seat 21 moves and makes each disposable capillary pipe 81 face the second joint 911 one by one, the pipetting table 4 descends, and the suction head assembly is pressed and raised.

The sliding seat 21 moves and the sample tube 15 is located below the second joint 911, the pipetting table 4 is lowered, and the vacuum pump 5 works to extract samples. The pipetting table 4 rises, the sliding seat 21 moves so that the first cleaning solution is located below the second joint 911, the pipetting table 4 descends, and the vacuum pump 5 works to extract the first cleaning solution. The pipetting table 4 rises, the sliding seat 21 moves so that the second cleaning liquid is located below the second joint 911, the pipetting table 4 is lowered, and the vacuum pump 5 works to extract the second cleaning liquid. The pipetting table 4 rises, the sliding seat 21 moves and the third cleaning liquid is located below the second joint 911, the pipetting table 4 is lowered, and the vacuum pump 5 works to extract the third cleaning liquid. The pipetting table 4 rises, the sliding seat 21 moves so that the fourth cleaning solution is located below the second joint 911, the pipetting table 4 descends, the vacuum pump 5 works, and the fourth cleaning solution is drawn and the filter element is vacuumed for 2-5 minutes.吸头83.

The pipetting table 4 rises, the sliding seat 21 moves and the first tip hole 71 is located below the second joint 911, the pipetting table 4 descends, the piston driving device 61 works, and drives the ejector plate 41 to retract the filter tip 83 To the first nozzle hole 71. The pipetting table 4 rises, the sliding seat 21 moves and the first reagent hole 73 is located below the second joint 911, the pipetting table 4 descends, the piston driving device 61 works, and the ejector plate 41 is driven to retract the disposable capillary pipe 81 To the first reagent hole 73 (the diluent in the first reagent hole has been evacuated, and thus is used to place the disposable capillary tube 81 after use). The pipetting table 4 rises, the sliding seat 21 moves and the filter tip 83 is located below the first joint 621a, the pipetting table 4 descends, and the filter tip 83 is pressed and raised. The sliding seat 21 moves and the third reagent hole 75 is located below the first joint 621a, the pipetting table 4 is lowered, and the piston driving device 61 operates to suck the eluent.

After the suction is completed, the pipetting table 4 rises, the sliding seat 21 moves and the fourth reagent hole 76 is located below the first joint 621a, the pipetting table 4 drops, and the piston driving device 61 works to push out the eluent.

The pipetting table 4 rises, the sliding motor 221 drives the sliding seat 21 so that the first suction head hole 71 is located below the first joint 621a, the pipetting table 4 descends, the piston driving device 61 operates and drives the ejector plate 41 to remove the filter element The suction head 83 retracts into the first suction head hole 71, and the motors are reset.

Compared with the prior art, the nucleic acid extraction device of the present invention can realize the automatic and rapid operation of nucleic acid extraction, and the whole process only takes 5 to 10 minutes, and the processing of large sample volume can not be realized in a short time, thereby realizing the efficient extraction of nucleic acid. In addition, the sample is purified by suction and cleaning, so that high-purity nucleic acid materials can be obtained without the risk of cross-contamination, with good repeatability and low cost.

Claims

A nucleic acid extraction device, comprising a workbench (1) with a countertop (10), characterized in that it further comprises:

A sliding seat (21), which is arranged on the table (10) and can slide back and forth along the table (10) under the drive of the sliding assembly (22);

The sample holder (3) is arranged on the sliding seat (21), and the sample holder (3) has a plurality of sample holes (31) arranged in parallel along the direction perpendicular to the sliding direction of the sliding seat (21) ), the sample hole (31) is used to place the sample tube (15);

The reagent strip holder (213), which is set on the sliding seat (21), is provided with a containing groove (213a) for placing the reagent strip (7), and the number of the containing grooves (213a) is equal to The number of the sample holes (31) matches, the reagent strips (7) all include a first tip hole (71), a second tip hole (72a, 72b) and a reagent hole arranged in a straight line, wherein , The first tip hole (71) is used to place the suction head assembly, the second tip hole (72a, 72b) is used to place the tip (82), the reagent hole includes the first reagent for containing the diluent Hole (73), the second reagent hole (74a, 74b, 74c, 74d) for containing the cleaning solution, the third reagent hole (75) for containing the eluent, and the second reagent hole (75) for storing the target substance Four reagent holes (76), each of the accommodating grooves (213a) contains the reagent strip (7), the first tip hole (71) on each reagent strip (7) ), the second tip holes (72a, 72b) and each of the reagent holes are directly opposite to the corresponding sample holes (31);

The pipetting table (4) is horizontally arranged on a stand (12) fixed on the table (10), and can be moved up and down relative to the sliding seat (21) under the driving of the lifting assembly (64);

The piston assembly, which is arranged on the pipetting table (4), includes a piston member (62) and a piston driving device (61), the number of the piston member (62) and the number of the sample holes (31) The numbers match, each of the piston parts (62) includes a piston tube (622) and a piston (621) built in the piston tube (622) and capable of moving up and down along the inner wall of the piston tube (622), and each The piston tubes (622) are vertically pierced on the pipetting table (4), and the lower ends of the piston tubes (622) are respectively formed with first joints (621a) for inserting and connecting the suction head (82) ), the piston driving device (61) can drive the piston (621) in each of the piston parts (62) to move up and down relative to the corresponding piston tube (622); and

The suction assembly includes a connecting suction pipe (91) and a vacuum pump (5). The number of the connection suction pipes (91) matches the number of the sample holes (31), and each of the connection suction pipes (91) is The upper end of each of the connecting pipettes (91) is respectively connected with the vacuum pump (5), and the lower ends are respectively formed with second joints for inserting and connecting the suction head assembly (911).
The nucleic acid extraction equipment according to claim 1, wherein a heating device for heating the sample holder (3) is further provided on the sliding seat (21).
The nucleic acid extraction device according to claim 1 or 2, further comprising a piercing assembly for piercing the seal on each reagent hole, the piercing assembly including a piercing rack (141) and The piercing head (14) on the piercing frame (141), the piercing head (14) is matched with the sample hole (31),

The pistons (621) are all mounted on a piston frame (63), the piston frame (63) is fixed on the output shaft of the piston driving device (61), and the piercing frame (141) and the piston frame ( 63) Fixed.
The nucleic acid extraction device according to claim 3, characterized in that there are multiple vacuum pumps (5) and one-to-one correspondence with the piston member (62), and a suction connector is provided on the piston frame (63) (92), the number of the suction joints (92) corresponds to the piston parts (62) one-to-one, and each suction joint (92) is threaded on the suction joint frame (65), and each suction joint (92) One end of the joint (92) is connected with the corresponding interface on the vacuum pump (5) through a connecting pipe, and the other end of the interface is connected with the upper end of the suction pipe (91) through a connecting pipe.
The nucleic acid extraction device according to claim 4, wherein a mounting frame (51) is provided on the table (10), and each vacuum pump (5) is placed on the mounting frame (51).
The nucleic acid extraction device according to claim 1 or 2, wherein the sliding component (22) comprises a first sliding block (227), a first sliding rail (226), a pressing block (225), a sliding The motor (221), the timing belt (224), the driving pulley (222) and the driven pulley (223), the first sliding block (227) is fixed at the bottom of the sliding seat (21), the first The sliding rail (226) is fixed on the table (10), the first sliding block (227) can slide back and forth along the first sliding rail (226), and the sliding motor (221) is fixed on the On the table (10), the driving pulley (222) is fixed on the output shaft of the sliding motor (221), and the driven pulley (223) is installed on the table (10). The timing belt (224) is tensioned between the driving pulley (222) and the driven pulley (223), and the pressure block (225) is fixed on one side of the sliding seat (21) and It is pressed on the timing belt (224).
The nucleic acid extraction device according to claim 1 or 2, wherein the lifting assembly (64) comprises a lifting motor (641), a lifting screw (642), and a lifting seat (643), and the stand (12) ) Includes a door frame (121) and a vertical plate (122) fixed on the top plate of the door frame (121), the lifting motor (641) is installed on the upper end of the vertical plate (122), and the vertical plate ( 122) A second sliding rail (122b) and a sliding hole (122a) extending vertically are respectively provided below the lifting motor (641), the lifting screw (642) is vertically arranged and its upper end is connected to the The output shaft of the lifting motor (641) is connected, and the lower end is connected with the top plate of the door frame (121), and the lifting seat (643) is penetrated on the vertical plate (122) through the sliding hole (122a) , And its front end is fixed to the pipetting table (4), and the rear end is sleeved on the lifting screw (642) and forms a screw transmission pair with the lifting screw (642), and the lifting seat (643) Is provided with a second slider (644),

When the lifting motor (641) drives the lifting screw (642) to rotate, the lifting screw (642) can drive the lifting seat (643) and enable the second sliding block (644) to move along the second The sliding rail (122b) moves up and down, and the corresponding part of the lifting seat (643) can move up and down along the sliding hole (122a).
The nucleic acid extraction device according to claim 1 or 2, wherein the reagent strip (7) comprises a carrier plate (70), and a plurality of first communication channels are arranged on the carrier plate (70) in parallel along its length. Hole (701) and a second through hole (702), one side of the carrier plate (70) is protruded with a tube body (703) corresponding to the first through hole (701), and the carrier plate ( 70) is also extended with a sleeve (704) corresponding to the second through hole (702) one-to-one, and the nozzle of each tube (703) is connected to the corresponding first through hole ( 701) and the openings of each sleeve (704) are respectively opposite to the corresponding second through holes (701), thereby respectively forming the first suction head hole (71) and the second suction head hole (72a) , 72b) and each reagent well.
The nucleic acid extraction device according to claim 8, wherein a first connector (78) and a second connector (77) are respectively provided on both sides of the carrier plate (70), and the first connector ( 78) and the second connecting member (77) can be connected in a detachable manner.
The nucleic acid extraction device according to claim 9, wherein the first connecting member (78) is a connecting post, and the second connecting member (77) is a connecting sleeve for inserting the connecting post.
The nucleic acid extraction device according to claim 1 or 2, wherein the suction head assembly comprises a filter tip (83) and a disposable capillary tube (81), and the filter tip (83) is embedded There is an adsorption filter element (830), the upper end of the disposable capillary straw (81) is sleeved on the lower end of the connecting straw (91) and embedded in the upper port of the filter element suction head (83), and The disposable capillary straw (81) is located in the filter tip (83) and above the adsorption filter core (830).
The nucleic acid extraction device according to claim 11, wherein the lower end of the disposable capillary pipette (81) is opposite to the adsorption filter element (830) up and down.
The nucleic acid extraction device according to claim 12, wherein the distance between the lower end of the disposable capillary pipette (81) and the upper end of the adsorption filter element (830) is 0-10 mm.
The nucleic acid extraction device according to claim 13, wherein the distance between the lower end of the disposable capillary pipette (81) and the upper surface of the adsorption filter element (830) is 2-5 mm.
The nucleic acid extraction device according to claim 11, wherein the disposable capillary pipette (81) comprises a first part (811) at the upper end and a second part (812) at the lower end, wherein the first part ( 811) is used to sleeve on the connecting straw (91), and the diameter of the second part (812) can allow the liquid to smoothly pass through the disposable capillary straw (81) under the action of suction.
The nucleic acid extraction device according to claim 15, wherein the pipe diameter of the first part (811) is 0.5-2.0 mm.
The nucleic acid extraction device according to claim 11, wherein the edge of the upper port of the disposable capillary pipette (81) is lower than the edge of the upper port of the filter tip (83).
The nucleic acid extraction device according to claim 11, wherein the inner peripheral surface of the upper end of the filter tip (83) is provided with a convex ring (831) along the circumferential direction, and the convex ring (831) can be connected to the primary The outer peripheral surface of the corresponding part of the sex capillary straw (81) is offset.
The nucleic acid extraction device according to claim 18, wherein the convex rings (831) are at least two arranged up and down, and each convex ring (831) is arranged up and down at intervals.